【摘要】 对布鲁氏菌疫苗靶标抗原铜锌超氧化物歧化酶(Cu/Zn SOD)蛋白进行生物信息学分析及重组表达。利用NCBI、Protter、Expasy、IEDB、SYFPEITHI、STRING数据库和SOPMA、Alpha-Fold软件等生物信息学工具对Cu/Zn SOD的序列保守性、理化性质、结构特点、表位分布及互作蛋白进行预测分析。采用全基因合成方法构建表达质粒pET-28a(+)-sodc,转化至BL21(DE3)感受态细胞中，应用IPTG诱导蛋白表达，并用SDS-PAGE和Western blot对蛋白进行鉴定，利用亲和层析柱对蛋白进行纯化。Cu/Zn SOD蛋白由173个氨基酸组成，理论相对分子量为18 130.67 u,等电点为6.24,N端第1位至第19位氨基酸为信号肽序列，在牛、羊、猪和犬种布鲁氏菌中高度保守，二级结构中α-螺旋、β-转角、无规则卷曲、延展结构的占比分别为9.83%、7.51%、57.23%和25.43%,存在多个T细胞和B细胞抗原表位并可与多种蛋白相互作用。此外，鉴定并纯化获得了Cu/Zn SOD重组蛋白。布鲁氏菌Cu/Zn SOD蛋白氨基酸序列保守性高、性质稳定、亲水性良好且涵盖了丰富的B/T细胞表位，可能有较好的免疫原性，为布鲁氏菌病新型疫苗的研发提供了参考。更多还原

【Abstract】 To perform bioinformatics analysis and recombinant expression of Cu/Zn superoxide dismutase(Cu/Zn SOD) protein, a target antigen for Brucellosis vaccines.A range of bioinformatics tools was adopted to predict and analyze the sequence conservation, physicochemical properties, structural features, epitope distribution, and interacting proteins of Cu/Zn SOD.The following tools were employed: NCBI,Protter, ExPASy, IEDB,SYFPEITHI,and STRING database, along with SOPMA and AlphaFold software.The expression plasmid pET-28a(+)-sodc was constructed using gene synthesis methods and transformed into BL21(DE3) competent cells.The plasmid was induced for expression using IPTG,and the protein was identified via SDS-PAGE and Western Blot.The protein was purified using an affinity chromatography column.The Cu/Zn SOD protein is comprised of 173 amino acids, with a theoretical molecular weight of 18 130.67 u and an isoelectric point of 6.24.The signal peptide sequence, which spans the first 19 amino acids at the N-terminus, exhibits high conservation among Brucella species in B.abortus,B.melitensis,B.suis,and B.canis.The secondary structure is comprised of 9.83% α-helix, 7.51% β-turns, 57.23% random coils, and 25.43% extended strand.It harbors multiple T-cell and B-cell antigenic epitopes and has the capability to interact with various proteins.Additionally, the Cu/Zn SOD was successfully constructed, identified, and purified.The Cu/Zn SOD protein of Brucella exhibits high amino acid sequence conservation, stability, and hydrophilicity.It also contains a broad range of B/T cell epitopes, which may enhance its immunogenicity, thereby providing a valuable reference for the development of Brucellosis vaccines.更多还原